| 鸡白细胞介素-15基因克隆、序列分析及表达 |
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| 引用本文： | 段丽娟,李明峰,宋晓琳,李凤华,孙宏鑫,江国托.鸡白细胞介素-15基因克隆、序列分析及表达[J].中国家禽,2006,28(22):17-19,22. |
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| 作者姓名： | 段丽娟 李明峰 宋晓琳 李凤华 孙宏鑫 江国托 |
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| 作者单位： | 1. 大连三仪生物工程研究所,辽宁,大连,116036
2. 大连轻工业学院生物与食品工程学院,辽宁,大连,116034 |
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| 摘 要： | 根据GenBank上发表的鸡IL-15基因序列设计两对引物,以从鸡脾脏中提取的基因组RNA为模板,采用RT-PCR扩增出鸡白细胞介素-15基因并进行了序列分析。序列分析结果表明,鸡白细胞介素-15基因序列全长580bp,开放阅读框架内564个核苷酸共编码188个氨基酸。所扩增的序列与已报道的序列同源性为99.9%(563/564)。将扩增序列插入大肠杆菌载体pBV220中进行表达,SDS-PAGE显示目的蛋白约为21ku,表达产物经免疫荧光鉴定为阳性。
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| 关 键 词： | 鸡白细胞介素-15 RT-PCR 克隆 表达 |
| 收稿时间： | 2006-06-12 |
| 修稿时间： | 2006-06-122006-07-05 |
Cloning,Sequencing and Expression of Chicken Interleukine-15 Gene |
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| Duan Lijuan,Li Mingfeng,Song Xiaolin,Li Fenghua,Sun Hongxin,Jiang Guotuo.Cloning,Sequencing and Expression of Chicken Interleukine-15 Gene[J].China Poultry,2006,28(22):17-19,22. |
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| Authors: | Duan Lijuan Li Mingfeng Song Xiaolin Li Fenghua Sun Hongxin Jiang Guotuo |
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| Institution: | 1.Dalian Sanyi Bioengineering Insititute, Dalian,Liaoning 116036; 2. Biology and Food Engineering,Dalian Institute of Light Industry,Dalian,Liaoning 116034 |
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| Abstract: | Based on the published nucleotide sequence of chicken Interleukine-15(IL-15)gene in GenBank,two pairs of primers were designed and synthesized. Total RNA,isolated from spleen of chicken,was used as template to generate complementary DNA by RT-PCR. Results demonstrated that chicken IL-15 gene contained 580 bp with open reading fragment(ORF)of 564 bp encoding a peptide of 188 amino acids. The amplified gene of IL-15 was 99.9%(563/564)homologous to the nucleotide acids of the Genbank. The cloned gene fragment was inserted into expression vector pBV220. The SDS-PAGE result showed that molecular weight of interest protein was about 21 ku. The expression was certified by immune-fluorescence technique. |
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| Keywords: | chicken interleukine-15 RT-PCR cloning expression |
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